Funds are requested to purchase an Illumina HiSeq 3000 Sequencer for the Genome Sequencing Facility (GSF) of Greehey Children's Cancer Research Institute (GCCRI) at the University of Texas Health Science Center at San Antonio (UTHSCSA). At the present time, high throughput sequencing is conducted on a heavily utilized Illumina HiSeq 2000 sequencer that was purchased in December 2010, and there were no other publicly accessible high-throughput NGS platforms within UTHSCSA and surrounding San Antonio institutions. Since the installing of the HiSeq 2000 at GCCRI GSF, it has quickly become essential for many research projects predominately funded by NIH. During the past four years, more than 6,000 samples from > 50 investigators of UTHSCSA and > 20 investigators from surrounding San Antonio areas and other institutions have been processed and sequenced at GSF, covering the wide-range sequencing protocols and applications. The heavy usage of the existing HiSeq 2000 has reached the capacity and facing some serious limitations including relative high cost and long running time of sequencing, comparing to the current model of HiSeq systems. Considering that the GSF's only HiSeq 2000 is not eligible for upgrade, we propose to purchase a much improved Illumina HiSeq 3000 sequencer with the trade-in option of HiSeq 2000 sequencer, such that GSF can continue provide the high quality genomic data generation to support NIH funded biomedical research as we demonstrated in the proposal. With significantly less cost, faster turn-around time, and higher data throughput we expect from the new sequencer, the GSF will continue our collaborative assistance to the development of genome-enabled research projects, scientific programs and grant proposals. In addition, we will continue our educational training programs to pre- and post-doctoral scientists, physicians, and scientists, and foster excellence of genomics and bioinformatics research at UTHSCSA. The projects of 16 major users and 13 minor users are presented. With the capacity of new requested Illumina HiSeq 3000 sequencer, a wide variety of medically relevant areas will be investigated including tumor heterogeneity identification, tumor subtype diagnosis and prognosis, cancer driver/rare mutation characterization, gene expression regulation and network of pathways, miRNA functions during early cancer detection, DNA damage, diabetes, and behavior science. In addition to the proposed user group, the new features of HiSeq system, such as lower cost, faster turnaround time, and longer sequence reads, will stimulate other investigators to plan new experiments in the future that may take their research endeavors into exciting new directions. Thus, acquisition of an illumina HiSeq 3000 has the potential to positively impact to a large number of NIH-funded investigations, further accelerate translational research to the patients in our unique dominant minority South Texas area.